LOCUS       Exported               14961 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pBLOXNPT, complete sequence.
ACCESSION   EU327975
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14961)
  AUTHORS   Ma BG, Ma C, Niu JX.
  TITLE     Gene expression controlled by inducible Cre/loxP recombination 
            system in transgenic tobacco
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14961)
  AUTHORS   Ma BG, Ma C, Niu JX.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-DEC-2007) Department of Horticulture, Agriculture 
            College Shihezi University, Shihezi, Xinjiang 832000, P.R. China
REFERENCE   3  (bases 1 to 14961)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14961
                     /organism="Binary vector pBLOXNPT"
                     /mol_type="other DNA"
                     /db_xref="taxon:490503"
     misc_feature    complement(13..796)
                     /label=similar to traF in GenBank Accession Number X54459
                     /note="similar to traF in GenBank Accession Number X54459"
     CDS             complement(145..516)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            549..658
                     /note="incP origin of transfer"
     rep_origin      complement(790..1168)
                     /direction=LEFT
                     /note="ColE1 ori; similar to sequence in GenBank Accession 
                     Number V00268"
     misc_feature    complement(1161..2344)
                     /label=similar to tetA in GenBank Accession Number X75761
                     /note="similar to tetA in GenBank Accession Number X75761"
     CDS             complement(1339..1989)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    complement(2454..2478)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     promoter        2511..2529
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2583..2616)
                     /label=loxP site
                     /note="loxP site"
     protein_bind    2583..2616
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2661..2967
                     /regulatory_class="promoter"
                     /note="NOS"
     promoter        2744..2923
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2776..2959
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2980..3774
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase II"
                     /label=nptII
                     /protein_id="ABY50072.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2980..3774
                     /gene="nptII"
                     /label=nptII
     regulatory      4164..4419
                     /regulatory_class="terminator"
                     /note="NOS"
     terminator      4164..4416
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(4695..4728)
                     /label=loxP site
                     /note="loxP site"
     protein_bind    4695..4728
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    5026..5047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5062..5092
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5100..5116
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5124..5140
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      5177..6011
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        5664..6009
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             6048..7859
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /note="GUS"
                     /protein_id="ABY50073.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            6048..7859
                     /gene="gusA"
                     /label=gusA
     regulatory      7930..8182
                     /regulatory_class="terminator"
                     /note="NOS"
     terminator      7930..8182
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8191..8207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(8824..8849)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    8825..8849
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(9359..10401)
                     /label=similar to tetA in GenBank Accession Number L13842
                     /note="similar to tetA in GenBank Accession Number L13842"
     misc_feature    complement(10402..11883)
                     /label=similar to trfA in GenBank Accession Number X00713
                     /note="similar to trfA in GenBank Accession Number X00713"
     CDS             complement(10445..11593)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    complement(11884..12876)
                     /note="similar to NPTIII gene in GenBank Accession Number 
                     V01547"
     CDS             complement(11892..12686)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    complement(12877..13646)
                     /note="similar to transposable element IS1 in GenBank 
                     Accession Number X58999"
     mobile_element  12878..13645
                     /mobile_element_type="insertion sequence:IS1"
                     /note="prokaryotic transposable element"
     misc_feature    complement(13647..13997)
                     /note="similarity to NPT III gene in GenBank Accession 
                     Number V01547"
     misc_feature    complement(13998..14269)
                     /label=similar to kilA in GenBank Accession Number M62846
                     /note="similar to kilA in GenBank Accession Number M62846"
     rep_origin      14268..18
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      complement(14344..14961)
                     /direction=LEFT
                     /note="ori V; similar to sequence in GenBank Accession 
                     Number M20134"